Immunocytochemistry

The cells were washed three times with DPBS (GIBCO) and fixed for 10 minutes with 4% paraformaldehyde (Merck Millipore), followed by three more rinses with DPBS.

The cells were blocked for 1h 0m 0s in blocking solution (KPBS 0.25% Triton X-100 (Fisher Scientific) and 5% normal donkey serum) at Room temperature.

The primary antibody was added to the blocking solution and incubated 1h 0m 0s at Room temperature (follow manufacturer's instructions for dilutions).

The cells were then washed the next day three times with KPBS 0.25%.

The secondary antibody was added with DAPI (1:1000; Sigma-Aldrich) to the blocking solution and incubated at Room temperature for 1h 0m 0s.

The cells were finally rinsed 2-3 times with KPBS and visualised on a Leica microscope (model DMI6000B).